caa a22 4500 477075134 CHVBK 20180405111437.0 cr unu---uuuuu 170330e19960301xx s 000 0 eng 10.1007/BF02827593 doi (NATIONALLICENCE)springer-10.1007/BF02827593 Expression of human cytomegalovirus DNA polymerase in insect cells using baculovirus expression system: Purification and biochemical characterizations [Elektronische Daten] [Ye Linbai, Gao Jinrong, Huang Engshang] Human Cytomegalovirus (HCMV) DNA polymerase gene was overexpressed in insect cells using the baculovirus transfer system. A6. 2 kb HCMV Rsr II-EcoRI DNA fragment with intact HCMV pol gene coding sequence was engineered into NheI site of vector pBlueBac under the control of polyhedrin promoter of Autographa californica nuclear polyhedrosis virus (AcNPV). Recombinant AcNPV carried HCMV pol gene was generated by cotransfection of Spodoptera frugiperta cell (SF21) with AcNPV DNA and baculovirus transfer vector with HCMV pol gene. Infection of SF21 cell with recombinant virus lead to the expression of 140 kD peptide of HCMV specific DNA polymerase at the level approximately 2 mg per 108 cells. The polypeptide was purified from the infected SF21 cells by a series of column chromatography to homogeneity. The purified enzyme had a molecular weight of 140 kD and reacted with antiserum specific for HCMV DNA polymerase. It exhibited both 3′-5′ and 5′-3′ exonuclease activities. This enzyme is also sensitive to phosphono acetate. Springer, 1996 DNA polymerase nationallicence expression nationallicence purifcation nationallicence characterization nationallicence Linbai Ye College of Life Sciences, Wuhan University, 430072, Wuhan, China aut Jinrong Gao College of Life Sciences, Wuhan University, 430072, Wuhan, China aut Engshang Huang Lineberger comprehensive Cancer Center, Department of Medicine, Microbiology and Immunilogy, and Curriculum of Genetics, University of North Carolona at Chapel Hill, 27699-7295, Chape Hill, NC aut Wuhan University Journal of Natural Sciences Wuhan University 1/1(1996-03-01), 107-115 1007-1202 1:1<107 1996 1 11859 https://doi.org/10.1007/BF02827593 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF02827593 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Linbai Ye College of Life Sciences, Wuhan University, 430072, Wuhan, China aut NATIONALLICENCE 700 1- Jinrong Gao College of Life Sciences, Wuhan University, 430072, Wuhan, China aut NATIONALLICENCE 700 1- Engshang Huang Lineberger comprehensive Cancer Center, Department of Medicine, Microbiology and Immunilogy, and Curriculum of Genetics, University of North Carolona at Chapel Hill, 27699-7295, Chape Hill, NC aut NATIONALLICENCE 773 0- Wuhan University Journal of Natural Sciences Wuhan University 1/1(1996-03-01), 107-115 1007-1202 1:1<107 1996 1 11859 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer